Abstract：
:An important area of proteomics involves the need for quantification, whether relative or absolute. Many methods now exist for relative quantification, but to support biomarker proteomics and systems biology, absolute quantification rather than relative quantification is required. Absolute quantification usually involves the concomitant mass spectrometric determination of signature proteotypic peptides and stable isotope-labeled analogs. However, the availability of standard labeled signature peptides in accurately known amounts is a limitation to the widespread adoption of this approach. We describe the design and synthesis of artificial QconCAT proteins that are concatamers of tryptic peptides for several proteins. This protocol details the methods for the design, expression, labeling, purification, characterization and use of the QconCATs in the absolute quantification of complex protein mixtures. The total time required to complete this protocol (from the receipt of the QconCAT expression plasmid to the absolute quantification of the set of proteins encoded by the QconCAT protein in an analyte sample) is approximately 29 d.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Pratt JM,Simpson DM,Doherty MK,Rivers J,Gaskell SJ,Beynon RJdoi
10.1038/nprot.2006.129subject
Has Abstractpub_date
2006-01-01 00:00:00pages
1029-43issue
2eissn
1754-2189issn
1750-2799pii
nprot.2006.129journal_volume
1pub_type
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